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Cucumber mosaic virus (CMV) is a virus that can infect various monocotyledonous and dicotyledonous plants. Two efficient detection methods for CMV in orchids, real-time quantitative PCR (RT-qPCR) and reverse transcription loop-mediated isothermal amplification (RT-LAMP) were developed in the study. For RT-qPCR, a TaqMan probe-based assay was designed using conserved regions of the coat protein (cp) gene, with a cloned cp plasmid serving as the standard for calibration curve construction. For RT-LAMP, specific inner and outer primers were designed based on the cp gene conserved sequences too. Both methods specificity detection were performed using virus RNA from CMV, Cymbidium mosaic virus (CymMV), and Odontoglossum ringspot virus (ORSV) as templates, and for sensitivity detection was performed using 10-fold serial dilutions of CMV RNA as a template. Additionally, field-collected orchid samples were screened for CMV infection using both techniques. The CMV RT-qPCR and RT-LAMP detection methods established in this study detected only CMV-positive samples without cross-reactivity with CymMV or ORSV. The sensitivity of RT qPCR and RT LAMP was consistent with a dilution of 106 times the original solution. The positive rate of CMV in field orchid samples was 26.7%. The results demonstrate that the RT-qPCR and RT-LAMP established in this study have strong specificity and high sensitivity, and are suitable for monitoring CMV infection in orchids in actual production processes.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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